[1]王彦,卓文磊,王长征,等.小鼠FasL全长cDNA的克隆、真核表达载体构建及其在树突状细胞中的表达[J].第三军医大学学报,2007,29(01):28-31.
 WANG Yan,ZHUO Wen-lei,WANG Chang-zheng,et al.Cloning and eukaryotic expression vector construction of mouse FasL gene and its expression in dendritic cells[J].J Third Mil Med Univ,2007,29(01):28-31.
点击复制

小鼠FasL全长cDNA的克隆、真核表达载体构建及其在树突状细胞中的表达(/HTML )
分享到:

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
29卷
期数:
2007年第01期
页码:
28-31
栏目:
论著
出版日期:
2007-01-15

文章信息/Info

Title:
Cloning and eukaryotic expression vector construction of mouse FasL gene and its expression in dendritic cells
作者:
王彦 卓文磊王长征钱桂生毕玉田吴奎
第三军医大学新桥医院:全军呼吸内科研究所,全军呼吸病研究重点实验室,全军肿瘤诊治中心,重庆市肿瘤生物技术研究所
Author(s):
WANG Yan ZHUO Wen-lei WANG Chang-zheng QIAN Gui-sheng BI Yu-tian WU Kui
Institute of Respiratory Diseases, Cancer Research Center, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China
关键词:
FasL 克隆 载体构建 树突状细胞
Keywords:
FasL clone vector construction dendritic cell
分类号:
Q782; Q785; Q786
文献标志码:
A
摘要:
目的     克隆小鼠FasL(mouse FasL, mFasL)全长cDNA,构建其真核表达载体,并验证其在小鼠骨髓来源的树突状细胞(dendritic cells, DC)中的表达。     方法     采用RT-PCR和TA克隆技术,从激活的小鼠脾脏T淋巴细胞中扩增mFasL全长cDNA,亚克隆到T载体中,再克隆到pIRES2EGFP载体中。转染小鼠DC后,用RT-PCR、免疫荧光及Western blot检测mFasL mRNA和蛋白表达。     结果     测序证实所得的mFasL cDNA序列与其在GenBank中的序列完全一致。mFasL真核表达载体转染小鼠DC后,能表达mFasL mRNA和蛋白。     结论     成功克隆了mFasL基因,构建其真核表达载体,并证明能有效表达于DC中。
Abstract:
Objective     To clone mouse FasL (mFasL) full length cDNA and construct eukaryotic expression vector, and to detect its expression in dendritic cells.      Methods     RT-PCR and TA cloning technique were used to clone the mFasL full length cDNA from mouse activated T lymphocytes, and then eukaryotic expression vector pIRES2EGFP containing mFasL cDNA was constructed. The positive  recombinants pIRES2EGFP-mFasL were transfected into dendritic cells (DC), the expression of mFasL mRNA was detected by RT-PCR and the expression of mFasL protein was assayed by immunofluorescence and Western blotting.      Results     The cloned full reading frame of mFasL cDNA was in coincidence with the sequence registered in GenBank. In the DC after transfection of pIRES2EGFP-mFasL, the expressions of mFasL mRNA and mFasL protein were detected.      Conclusion     The mFasL cDNA was cloned and constructed into the eukaryotic expression vector successfully, and mFasL gene could be expressed efficiently in DC transfected by pIRES2EGFP-mFasL.

参考文献/References:

王彦, 卓文磊, 王长征, 等. 小鼠FasL全长cDNA的克隆、真核表达载体构建及其在树突状细胞中的表达[J]. 第三军医大学学报,2007,29(1):28-31.

相似文献/References:

[1]曹唯希,王亚平,刘琼,等.人参皂甙诱导白血病细胞凋亡及对Fas、FasL表达的影响[J].第三军医大学学报,2005,27(06):525.
[2]刘青松,朱兴春,邢艳,等.丹参诱导类风湿关节炎患者成纤维样滑膜细胞中Fas/FasL的表达[J].第三军医大学学报,2010,32(22):2452.
[3]陈杰,辛海明,蔡云,等.人iASPP全长CDS的分段扩增、克隆及鉴定[J].第三军医大学学报,2007,29(11):996.
 CHEN Jie,XIN Hai-ming,CAI Yun,et al.Cloning and identification of human iASPP full-length CDS region[J].J Third Mil Med Univ,2007,29(01):996.
[4]吴国英,靳风烁,李黔生,等.Fas、FasL在肾癌组织中的表达及其意义[J].第三军医大学学报,2004,26(16):0.[doi:10.16016/j.1000-5404.2004.16.026 ]
 WU Guo ying,JIN Feng shuo,LI Qian sheng,et al.[J].J Third Mil Med Univ,2004,26(01):0.[doi:10.16016/j.1000-5404.2004.16.026 ]
[5]马铮,王如文,蒋耀光,等.重症肌无力胸腺细胞凋亡与Fas基因表达的研究[J].第三军医大学学报,2004,26(17):0.[doi:10.16016/j.1000-5404.2004.17.031 ]
 MA Zheng,WANG Ru wen,JIANG Yao guang,et al.[J].J Third Mil Med Univ,2004,26(01):0.[doi:10.16016/j.1000-5404.2004.17.031 ]
[6]龙冬梅,高宁.细胞凋亡相关基因Fas和FasL在大鼠苯巴比妥促肝癌过程中的表达研究[J].第三军医大学学报,2003,25(21):0.[doi:10.16016/j.1000-5404.2003.21.037 ]

更新日期/Last Update: 2008-11-13